Composition for diminishing neutral fat in blood

ABSTRACT

The present invention provides a neutral fat reducing composition which reduces a blood neutral fat safely and efficiently, and whose active ingredient is a soybean protein containing 50% or more of a 7S globulin fractionated from a soybean protein and 0.2% or less of phytate based on the proteins. Its effect is further enhanced by removing a oil-body-associated protein and reducing a chloroform methanol-extractable oil fraction.

FIELD OF THE INVENTION

The present invention relates to a blood neutral fat reducingcomposition.

PRIOR ART

Among vegetable proteins, a soybean protein not only has an excellentnutritional property but also was found recently to have variousphysiological efficacies and became an attractive food material as aphysiologically-functional agent.

A blood neutral fat reducing effect of a soybean protein has alreadybeen established based on a body fat-reducing effect of the soybeanprotein, and its mechanism was reported by Iritani, et al., (J. Nutr.126, 380, 1996) to be an inhibitory effect on the activity of a fattyacid synthetase in a liver. In addition, each of a whole soybeanglobulin, a 7S globulin and a 11S globulin was examined for its effecton fats in blood and a liver, and was reported generally to be moreexcellent in terms of an ability of reducing blood cholesterol orneutral fat when compared with casein which is an animal protein (Okitaet al., J. Nutr., 27, 379, 1981).

A 11S globulin-defect soybean, i.e., a 7S globulin-rich seed, which wasobtained by a breeding, was also reported (Breeding Science, 46, 11,1996) together with its utility (Breeding Science, 50, 101, 2000) andits patent (U.S. Pat. No. 6,171,640 B1).

Nevertheless, a soybean protein, including a 7S globulin, is known toform a complex with phytate by which the digestion of the soybeanprotein is affected adversely (M. A. Ritter, et al., J. Food Sci., 52,325, 1987).

As soybean-derived proteins, proteins having high affinities with polarlipids as constituents of a cytoplasmic membrane as well as protein bodyor oil body membrane have also been identified and designated by Samotoet al. as “Oil-Body-Associated Proteins”. An oil-body associated proteinis a general term of the proteins consisting mainly of membraneproteins, especially those whose molecular weights measured by anSDS-polyacrylamide gel electrophoresis were 34 kDa, 24 kDa and 18 kDa,and exist as a fraction containing about 10 to 12% by weight of polarlipids which are extractable with a 2:1 polar solvent mixture ofchloroform:ethanol, and were reported by Samoto et al. to be produced inan amount as high as about 35% of industrially produced fractionatedsoybean proteins (B.B.B., 62 (5), 935–940 (1998)). Anoil-body-associated protein has a poor flavor and a high allergenicity.

Nevertheless, since an oil-body-associated protein is stained onlyslightly in an SDS-polyacrylamide gel electrophoresis employedfrequently for determining the composition of a soybean protein, itsestimated amount becomes far less than its actual amount or almostnegligible. Thus, a conventional fractionation is focusing only on a 7Sand an 11S, and pays no attention to the oil-body-associated proteinswhich are contaminating each fraction. However, from a physiologicalpoint of view, the behavior of these oil-body-associated proteins shouldbe taken into account for the purpose of obtaining the 7S and the 11S athigher purities.

To prepare a 7S globulin efficiently from a soybean protein whilecleaving phytate binding as a complex off and suppressing thecontamination with the oil-body-associated proteins whereby raising thepurity is very important in the use not only as an extremely safepharmaceutical material for reducing the blood neutral fat but also as afood material.

OBJECTS OF THE INVENTION

An objective of the invention is to obtain a fraction having a bloodneutral fat reducing ability from the soybean proteins and to treat thisfraction for enhancing its ability, whereby providing it as a food or apharmaceutical.

SUMMARY OF THE INVENTION

We studied intensively and obtained the following findings.

(1) When removing an 11S globulin by a method by Thahn and Shibasakiwhich is a standard procedure for fractionating the soybean proteinsfrom a defatted soybean followed by fractionating a 7S globulintherefrom, a fractionation at a purity as high as 50% or higher ispossible without using any reducing agent.

(2) When the 7S globulin described above and a 7S globulin obtainedafter cleaving phytate binding to the 7S globulin off (hereinafterreferred to as a phytate-reduced 7S globulin) were subjected to a studyin rats for 21 days using casein as a reference control, the 7S globulinand the phytate-reduced 7S globulin exhibited more excellent bloodneutral fat reducing abilities when compared with the casein, with thephytate-reduced 7S globulin exhibiting an especially high neutral fatreducing effect.

(3) In response to the onset of this neutral fat reducing effect, theblood cholesterol level is reduced. While this cholesterol reducingability is exhibited by both of the 7S globulin and the phytate-reduced7S globulin, the HDL-cholesterol (HDLC) level is improved rather by thephytate-reduced 7S globulin.

The HDLC is so-called good cholesterol which recovers excessivecholesterols, and an improved HDLC leads to a great reduction in thearterial sclerosis index when combined with a total cholesterol reducingeffect.Arterial sclerosis index=(total cholesterol−HDL cholesterol)/HDLcholesterol

(4) By further treating the soybean proteins with a phytate decomposingenzyme, a low phytin 7S globulin from which phytate has been cleaved offcan be fractionated.

(5) By further removing a membrane protein rich oil-body-associatedprotein which contaminates a soybean protein, the efficacy of the 7Sglobulin as an active ingredient can further be enhanced.

As a result, it was discovered that by cleaving phytate off from a 7Sglobulin considered to have a blood neutral fat reducing effect and byremoving oil-body-associated proteins to raise the purity a higherefficacy can be achieved together with a corresponding reduction in thedose, whereby establishing the present invention.

The invention provides a blood neutral fat reducing composition whoseactive ingredients are 7S globulin-rich phytate-reduced soybeanproteins. Also provided is a blood neutral fat reducing compositionwhose active ingredient is a phytate-reduced highly purified 7S globulinwhich is a 7S globulin-rich soybean protein obtained by purifying afraction containing a large amount of a 7S globulin as a majorconstituent of the soybean proteins to a protein purity (on the basis ofSPE described below) as high as 50% or higher, in which thechloroform:methanol-extractable polar lipids as the indexes of theoil-body-associated protein have been reduced to a level of ⅓ or lessand in which the phytate content, which is about 2% based on theproteins in a commercial soybean protein product, has been reduced to alevel of 0.2% or less based on the proteins by cleaving phytate off.Thus, by means of a fractionation involving a suppression of thecontamination with the membrane protein rich oil-body-associatedproteins as far as possible and a cleavage of phytate, a blood neutralfat reducing composition having a further enhanced efficacy can beprovided. On the other hand, by employing a phytate decomposing enzymeduring the manufacturing process of a soybean protein, the fractionationof a 7S globulin is facilitated, whereby providing a blood neutral fatreducing composition containing as an active ingredient a highlypurified phytate-reduced 7S globulin resulting from a reduction inphytate to 0.2% or less based on the proteins and from a reduction inthe oil-body-associated proteins to 10% or less. Also provided is ablood neutral fat reducing composition produced without using anyreducing agents during the manufacturing process described above.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

In this invention, a 7S globulin means a globulin whoseultracentrifugation sedimentation coefficient is 7S among the solubleglobular proteins referred to generally as globulins. Based on themolecular weight distribution, the globulins are classified into 2S, 7S,11S and 15S, among which 7S and 11S are known to be abundant in thedepot proteins of a pulse plant such as a soybean. The 7S globulin of asoybean is substantially the same to β-conglycinin which is animmunological term.

In this invention, a 7S globulin-rich fraction obtained from soybeanproteins is employed as a main ingredient. The first step forfractionating the 7S globulin from the soybean proteins involves aremoval of an 11S globulin. This removal can be effected by a method byThahn and Shibasaki (Thahn, V. H., and Shibasaki, K., J.Agric.FoodChem., 24, 117, 1976) which is employed widely in these days forobtaining each globulin fraction, as well as a cold insoluble fraction(CIF) method utilizing a cryoprecipitation (Briggs, D. R., and Mann, R.L., Cereal Chem, 27, 243, 1950) and a fractionation with adding 0.1 Ncalcium chloride proposed by Wolf et al (Wolf, W. J., and Sly, D. A.,Cereal Chem, 44, 653, 1967). A fraction made free of the 11S and theoil-body-associated proteins by the method described below in Productionexample 2 may also be employed.

After removing the 11S globulin by any of the methods described above, a7S globulin is fractionated by an ordinary method for preparing anisolated soybean protein. In such procedure, a 7S globulin having apurity which is sufficiently acceptable for use can be obtained in theabsence of a reducing agent, and such absence of the reducing agent isfavorable for a wider application also when being used as a neutral fatreducing agent. A 7S globulin-rich soybean protein whose protein purity(on the basis of SPE) is 50% or higher, preferably 60% or higher, morepreferably 80% or higher, particularly 85% or higher can be obtained.The 7S globulin-rich fraction thus obtained is further subjected to atreatment with an enzyme having a phytate decomposing activity such asphytase or phosphatase or a formulation thereof, whereby cleavingphytate off. As a result, the blood neutral fat reducing abilitypossessed naturally by a phytate-reduced 7S globulin obtained bycleaving phytate to a level of 0.2% or less, preferably 0.1% or less,more preferably 0.05% or less based on the proteins is enhanced.

A method for fractionating the phytate-reduced 7S globulin obtained bycleaving phytate as described above may be accomplished simultaneouslywith a removal of a 11S globulin by treating a soybean protein directlywith an enzyme having a phytate decomposing activity such as phytase andphosphatase as well as a formulation thereof.

Further by removing the oil-body-associated proteins, the blood neutralfat reducing ability of the phytate-reduced 7S globulin can further byenhanced. For this purpose, the oil-body-associated proteins can beremoved as a precipitation at pH 5.6 to 6.8 after heating (30 to 75° C.)at a weakly acidic pH (pH 3.8 to 6.8) at which they tend to beinsoluble.

A composition of the invention can be formulated as an oral compositionwhose active ingredient is a fraction obtained as described above or asoybean protein, and formulated into various dosage forms such aspowders, sugar-coated tablets and granules by known methods optionallytogether with other excipients and additives.

A fraction or a soybean protein employed as an active ingredientaccording to the invention is a safe edible material the amount of whichto be incorporated into a composition or to be ingested is not limitedparticularly, and may be ingested as it is or may be incorporated into afood product for a dietary therapy. A preferable amount to be ingestedper kg body weight is 50 to 500 mg, preferably 100 to 300 mg as 7Sglobulin-rich phytate-reduced soybean proteins.

EXAMPLES

The usefulness of the present invention is further discussed in thefollowing examples, which are not intended to restrict the technicalspirit of the invention.

Major analytical methods employed in the invention are described below.

-   Crude protein; Based on a Kjeldahl method, a nitrogen content was    determined and multiplied by the coefficient 6.25 to convert into a    crude protein.-   SDS-Polyacrylamide electrophoresis; Based on a method by Laemmli    (Nature, 227, 680 (1970)), an analysis was conducted with a gradient    gel concentration from 10 to 20%. The amount of a sample applied was    10 μg.-   Phytate; A method by Alii Mohamed (Cereal Chemistry 63,    475–478, 1986) was employed.-   Chloroform methanol-extractable oil fraction; A dried sample was    combined with an about 50-fold volume of a mixture of chloroform and    methanol (2:1, v/v) and the weight ratio of the solids extracted by    reflux was determined as a chloroform methanol-extractable oil    portion.-   Purity (SPE standard); The pattern of the bands obtained in the    SDS-polyacrylamide electrophoresis described above was measured by a    densitometer and a % area of the corresponding band based on the    total area was represented as a purity (on basis of SPE). A 7S    globulin content mentioned here means the total amount of α, α′ and    β subunits, while an 11S globulin content means the total amount of    acidic polypeptides (A) and basic polypeptides (B).-   Corrected purity; Based on a purity (on basis of SPE) obtained above    and taking any contaminating oil-body-associated proteins, a    corrected purity was calculated as described below. Thus, a purity    of a sample (on basis of SPE) is represented as A %, and a purity is    calculated as a value based on the total proteins including the 7S    globulin, the 11S globulins and the oil-body-associated proteins,    since the sample contains the oil-body-associated proteins in an    amount at least 10 times the chloroform-methanol-extractable oil    portion in addition to the 7S globulin and the 11 S globulin.-   Corrected purity (%)=(100 (%)−chloroform·methanol-extractable oil    portion (%)*10)*A (%)/100-   Neutral fats, total cholesterols, HLD cholesterols (HDLC); A DRYCHEM    model 5500 manufactured by FUJI FILM was employed in a solid phase    method.

Preferred embodiments of the invention are described below.

Production Example 1 Preparation of 7S Globulin=“7S” and Phytate-Reduced7S Globulin=“7S-PH”)

A defatted soybean was combined with water at a weight ratio of 1:10 andstirred with adjusting at pH 7.0 for 1 hour and then centrifuged (4000r.p.m, 20° C., 10 minutes) to remove a precipitate. The resultantsupernatant was adjusted at pH 6.4, allowed to stand at 4° C. overnightand centrifuged (4000 rpm, 4° C., 10 minutes) to remove the precipitate.The resultant supernatant was adjusted at pH 4.5 and centrifuged (4000rpm, 4° C., 10 minutes), and the resultant precipitate was recovered asa 7S globulin curd.

This 7S globulin curd was combined with a 4-fold volume of water,adjusted at pH 6.0, supplemented with phytase (NOVO, PHYTASE NOVO L) at0.2% based on the proteins, and then allowed to react at 40° C. for 1hour. The reaction mixture was adjusted at pH 5.0, centrifuged (4000r.p.m., 20° C., 10 minutes) to remove a whey fraction, whereby obtaininga phytate-reduced 7S globulin curd. The both of the 7S globulin curd andthe phytate-reduced 7S globulin curd were hydrated and then neutralizedat pH 7.0, sterilized at 140° C. for 15 seconds, and then rapidlyspray-dried to obtain a 7S globulin and a phytate-reduced 7S globulin.Each of the 7S globulin and the phytate-reduced 7S globulin thusobtained was subjected to a SDS-polyacrylamide gel electrophoresis, andthe intensity of the color of a stained protein band revealed that thepurity was 80%. The phytate contents in the both were 1.8% and 0.05%,respectively, revealing that the phytase treatment resulted in an almostcomplete cleavage of the phytate. The chloroform methanol-extractableoil portion of this sample was 2.8%. On the other hand, the total amountof the sulfur-containing amino acids cystine and methionine was 25 mg/gprotein, which was higher than 5 mg/g protein exhibited naturally by apurified 7S, suggesting that a substantial amount of impurities stillremained.

Production Example 2 Preparation of Highly Purified Phytate-Reduced 7SGlobulin:“7S-PH-LP”)

One part by weight of a low-modified defatted soybean was combined with10 parts by weight of extraction water at 40° C., and adjusted at pH 5.3with hydrochloric acid. This solution was supplemented with 8 units perprotein of phytase (NOVO, PHYTASE NOVO L) and subjected to a proteinextraction simultaneously with an enzyme reaction for 30 minutes at 40°C. to obtain an enzyme-treated slurry extract. This enzyme-treatedslurry extract was cooled to about 25° C., adjusted at pH 6.1 withhydrochloric acid, and centrifuged (3000G) using a batch-typecentrifuge. Upon this, a discrete separation between a soluble fractionand an insoluble fraction was observed. The solution temperature of thiscentrifuge was about 25° C. Subsequently, the soluble fraction wasadjusted at pH 4.9 with hydrochloric acid, and centrifuged to obtain aprecipitate curd. The precipitate curd was washed with a 10-fold volumeof water, hydrated (to 4-fold weight), neutralized with sodiumhydroxide, sterilized at 140° C. for 15 seconds, and then rapidlyspray-dried to obtain a phytase-treated 7S globulin-rich fractionatedsoybean protein (“7S-PH-LP”).

The phytate-reduced 7S globulin thus obtained was subjected to anSDS-polyacrylamide gel electrophoresis and the intensity of the color ofa stained protein band revealed that the purity was 95%. The phytatecontent was 0.05% based on the proteins, indicating almost completecleavage of the phytate. On the other hand, thechloroform.methanol-extractable oil portion of this sample was 0.5%,which was substantially lower when compared with production example 1.The total amount of the sulfur-containing amino acids cystine andmethionine was 12 mg/g protein, suggesting that a highly purified 7S wasobtained whose impurities were substantially low in view of the value 5mg/g protein exhibited naturally by a purified 7S.

Example 1 Verification of Blood Neutral Fat Reducing Effect in Rats

A blood neutral fat reducing effect was verified in rats. Each feedemployed here contained as a dietary protein the soybean proteinobtained in production example 2 described above or casein (vitamin-freecasein, ORIENTAL YEAST) as a control at the concentration of 20%,together with 0.5% of cholesterol and 0.125% of sodium cholate as wellas a 1:2 mixture of sucrose and corn starch. A typical formulation isrepresented in table 1 shown below.

TABLE 1 Feed formulation of each treatment group Component Composition(%) Protein Adjusted at 20% as crude protein with α-corn starch Sucrose20.0 Corn oil 5.0 Vitamin mix 1.0 Mineral mix 3.5 Powdered cellulose 5.0Choline hydrogen tartarate 0.2 Cholesterol 0.5 Sodium cholate 0.125α-Corn starch to 100 in total

Experimental animals were 5-week old (growing period) and 20-week old(mature period) male Wistar rats (weighing from 100 to 120 g, and 330 to360 g) purchased from NIPPON SLC, and received a commercial solid chow(ORIENTAL YEAST, CRF-1) preliminarily for 1 week, and then divided intotwo treatment groups in total each consisting of 6 animals with nodeviation in the body weight between groups, and the animals were raisedfor 10 days with test feeds. Each rat was housed in an individual cageat a temperature of 23±1° C. and a humidity of 55±5% under a 12-hourlighting period (7:00 am to 7:00 pm). During the raising period, theanimals were allowed to receive water and feed ad libitum.

The treatment period was 10 days, during which the body weight wasmonitored. The results are represented in tables 2 and 3 shown below. Nosignificant difference in the weight gain was observed between thegroups.

TABLE 2 Change in body weight in treatment group of 5-week old rats,unit: g Phytate-reduced 7S Treatment group Casein globulin (7S-PH)Initiation of treatment 153.3 ± 1.3 153.3 ± 1.3 Termination of 201.7 ±1.3 192.0 ± 1.5 treatment Weight gain  48.4 ± 1.3  38.7 ± 1.6

TABLE 3 Change in body weight in treatment group of 20-week old rats,unit: g Phytate-reduced 7S Treatment group Casein globulin (7S-PH)Initiation of treatment 357.2 ± 2.7 356.4 ± 5.8  Termination of 358.7 ±3.7 352.0 ± 10.3 treatment Weight gain  0.3 ± 2.1  4.4 ± 4.6

After the 10-day treatment period, each animal was fasted for 7 hoursfrom the morning (8:00 am) on the 11th day, and then subjected to alaparotomy under an anesthesia with nembual, and a blood was taken froman abdominal aorta via a heparinized syringe. The blood was centrifuged(3000 r.p.m., 5° C. for 15 minutes) to separate a plasma, which wasexamined for the neutral fats and the cholesterols. The mean and thestandard deviation of the data in each group were calculated, and astatistical significance was analyzed using Duncan's multiple rangetest. The results are shown in tables 4 and 5. In the tables, a %neutral fat reduction and a % cholesterol reduction are the ratios (%)of the difference in respective data between the casein group and thetreatment group based on the data in the casein group.

TABLE 4 Change in blood neutral fats in treatment group of 5-week oldrats Phytate-reduced 7S Treatment group Casein globulin (7S-PH) Neutralfat level 176.2 ± 10.9 89.1 ± 5.7 % Neutral fat reduction — 49.6 Totalcholesterol level 144.1 ± 6.0  93.5 ± 2.9 % Cholesterol — 35.1 reductionUnit; (Level) mg/dl, (ratio) %

TABLE 5 Change in blood neutral fats in treatment group of 20-week oldrats Phytate-reduced 7S Treatment group Casein globulin (7S-PH) Neutralfat level 261.2 ± 22.5 108.6 ± 21.6 % Neutral fat reduction — 58.4 Totalcholesterol level 132.5 ± 3.5   92.8 ± 14.5 % Cholesterol — 30.0reduction Unit; (Level) mg/dl, (ratio) %

As evident from the data shown above, the phytate-reduced 7S globulinexhibited marked cholesterol- and blood neutral fat-reducing effects inboth of the growing period (5-week old) and the mature period (20-weekold).

Comparative Example 1 Verification of Blood Neutral Fat Reducing Effectin Rats

Similarly to example 1, a blood neutral fat reducing effect was verifiedin rats. A treatment chow had the composition similar to that in example1, except for using as a soybean protein the 7S globulin (7S) obtainedin production example 1.

Experimental animals were 5-week old male Wistar rats (weighing from 90to 10 g) purchased from NIPPON SLC, and received a commercial solid chow(ORIENTAL YEAST, CRF-1) preliminarily for 1 week, and then divided intotwo treatment groups in total each consisting of 6 animals with nodeviation in the body weight between groups, and the animals were raisedfor 3 weeks with test feeds. Each rat was housed in an individual cageat a temperature of 23±1° C. and a humidity of 55±5% under a 12-hourlighting period (7:00 am to 7:00 pm). During the raising period, theanimals were allowed to receive water and feed ad libitum.

The treatment period was 21 days, during which the body weight wasmonitored. The results are represented in table 6 shown below. Nosignificant difference in the weight gain was observed between thegroups.

TABLE 6 Change in weight in treatment group Unit: g Treatment groupCasein 7 globulin (7S-PH) Initiation of treatment 151.0 ± 1.3 151.1 ±0.9 Termination of 243.8 ± 1.6 240.8 ± 5.5 treatment Weight gain  92.7 ±1.5  89.7 ± 6.0

After the 21-day treatment period, each animal was fasted for 6 hoursfrom the morning (8:00 am) on the 22nd day, and then subjected to alaparotomy under an anesthesia with nenbual, and a blood was taken froman abdominal aorta via a heparinized syringe. The blood was centrifuged(3000 r.p.m., 5° C. for 10 minutes) to separate a plasma, which wasexamined for the neutral fats and the cholesterols. The mean and thestandard deviation of the data in each group were calculated, and astatistical significance was analyzed using Duncan's multiple rangetest. The results are shown in table 7. Each % reduction was determinedby a method similar to those for tables 4 and 5 in example 1.

TABLE 7 Change in blood neutral fats in treatment group Unit: gTreatment group Casein 7S globulin (7S-PH) Neutral fat level 190.4 ±12.2 122.7 ± 6.6 % Neutral red — 35.6 reduction Total cholesterol level118.7 ± 4.5  109.9 ± 5.8 % Cholesterol —  7.4 reduction Unit; (Level)mg/dl, (ratio) %

When compared with the cholesterol- and neutral fat-reducing effects ofthe phytate-reduced 7S globulin employed in example 1 relative tocasein, the reducing effect of the 7S globulin in comparative example 1was lower, revealing that the phytate-reduced 7S globulin had a higherblood fat improving effect when compared with the 7S globulin.

Example 2

The blood neutral fat reducing effect of the highly purifiedphytate-reduced 7S globulin (7S-PH-LP) prepared in production example 2was investigated. As controls, the casein employed in example 1, the 7Sglobulin (7S), the phytate-reduced 7S globulin (7S-PH) and a commercialseparated soybean protein (SPI) were employed. The composition of eachprotein is shown in table 8.

TABLE 8 Unit: % SPI 7S 7S-PH 7S-PH-LP Crude protein 86.2 88.0 90.5 92.3Purity (on basis of SPE) — 80.0 82.2 95.7 Chloroform · methanol- 3.2 2.82.8 0.5 extractable oil fraction Phytate 1.8 1.8 0.05 0.05 Correctedpurity — 57.6 59.2 90.9

5-Week old rats similar to those in example 1 were employed asexperimental animals and raised for 2 weeks during which the body weightwas monitored under the conditions similar to those in example 1. Afterthe treatment period, a blood was taken similarly to example 1, andexamined for the neutral fats and the cholesterols.

The results included the change in the body weight shown in table 9, andthe neutral fats, the cholesterol, HDLC and the arterial sclerosis indexcalculated therefrom shown in table 10.

TABLE 9 Unit; g Casein SPI 7S 7S-PH 7S-PH-LP Initiation of 138.5 138.7135.8 135.5 137.8 treatment Termination of 210.2 212.9 206.9 179.6 206.5treatment Weight gain 71.7 74.3 71.1 44.0 68.8

TABLE 10 Unit; mg/dl Casein SPI 7S 7S-PH 7S-PH-LP Neutral fat level178.3 120.0 105.0 92.7 91.0 Total cholesterol 125.6 88.3 88.3 90.8 87.3level HDLC 37.1 36.8 34.2 43.5 44.0 Arterial sclerosis 2.39 1.40 1.581.09 0.98 index

Based on the results described above, the neutral fat- and thecholesterol-reducing effects were higher in the order shown below,Casein<<SPI=7S<7S-PH<7S-PH-LP

and HDLC was revealed to be improved in a phytate-free group. Thus, thehighly purified phytate-reduced 7S globulin obtained by removing phytateand also by removing the oil-body-associated proteins exhibited the mostevident serum lipid improving effect, especially an arterial sclerosisindex reducing effect.

INDUSTRIAL APPLICABILITY

The present invention enables an efficient reduction in the bloodneutral fat level, and an inventive composition, which is highly safe,is not only useful as a prophylactic or therapeutic agent for thepurpose of reducing the blood neutral fat level but also capable ofserving as a food product for such purpose.

1. A composition for reducing the blood level of neutral fats,comprising as an active ingredient a 7S globulin-rich phytate-reducedsoybean protein, whose phytate content is 0.2% or less based on thesoybean protein, wherein the composition is capable of reducing theblood level of neutral fats.
 2. The composition according to claim 1comprising 10% or less of an oil-body-associated protein based on thesoybean protein.
 3. The composition according to claim 1 comprising 1%or less of a chloroform:methanol (2:1)-extractable oil portion base onthe soybean protein.
 4. The composition according to claim 1 which isobtained by reacting a soybean protein with a phytate decomposingenzyme.
 5. The composition according to claim 1 which is preparedwithout using any reducing agents over its manufacturing process.
 6. Amethod of reducing the blood level of neutral fats, comprisingadministering the composition of claim 1 to a subject in need ofreducing the blood level of neutral fats.